Differential RNA fingerprinting as a tool in the analysis of spermatozoal gene expression

Miller, D. H.; Tang, Pei-Zhong; Skinner, Clare; Lilford, Richard

doi:10.1093/oxfordjournals.humrep.a138607

Cited by 86 publications

(51 citation statements)

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“…The purity of these samples was estimated by RT-PCR using protamine-2 specific primers [13] that produce a 310 bp in samples containing genomic DNA (not shown). The reverse-transcribed cDNA amplicon is 148 bp long.…”

Section: Patients and Rna Samplesmentioning

confidence: 99%

Sperm transcriptome profiling in oligozoospermia

Montjean¹,

Grange²,

Gentien

et al. 2011

J Assist Reprod Genet

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Purpose Investigate in what extent sperm transcriptome of infertile men is different from that of fertile individuals. Methods Semen samples were collected for determination of sperm parameters as well as for RNA isolation. Gene expression profile was investigated in spermatozoa of 8 infertile and 3 fertile men by microarray analysis using the Affymetrix Chip HG-U133 Plus 2.0. Result(s)We observed up to 33-fold reduction expression of genes involved in spermatogenesis and sperm motility. Furthermore, there is an important decrease in expression of genes involved in DNA repair as well as oxidative stress regulation. In this study, we also show a striking drop in expression of histone modification genes. Conclusion(s) We found that transcription profile in germ cells of men with idiopathic infertility is different from that of fertile individuals. Interestingly, about 15% of the regulated genes (Eddy Rev Reprod 4:23-30, 1999) play a role in spermatogenesis.

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Section: Patients and Rna Samplesmentioning

confidence: 99%

Sperm transcriptome profiling in oligozoospermia

Montjean¹,

Grange²,

Gentien

et al. 2011

J Assist Reprod Genet

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“…RNA identification in mature sperm has been performed using techniques based on the detection of RNA by means of PCR after reverse transcription [59]. Our study investigated the expression of the FasL gene in ejaculated sperm and demonstrated that expression of the FasL gene was higher in adolescents with varicocele.…”

Section: Discussionmentioning

confidence: 95%

Expression of the Fas-ligand gene in ejaculated sperm from adolescents with and without varicocele

Giudice

Lima

Cenedeze

et al. 2010

J Assist Reprod Genet

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Purpose To assess FasL mRNA levels in ejaculated sperm from adolescent patients with and without varicocele. Methods Semen was obtained by masturbation following 2-4 days of ejaculatory abstinence, from 14 adolescents with varicocele grades II and III (study group), and 20 adolescents without varicocele (control group). Seminal analysis was done according to World Health Organization guidelines and morphology using Kruger's strict criteria. The Fas-ligand (FasL) gene expression was performed using reverse transcription and real-time quantitative polymerase chain reaction (RQ-PCR) analysis, according to the expression level of the housekeeping cyclophilin A gene. A Student's t-test was applied to compare the groups, and Spearman's rank test in order to verify possible correlations (p<0.05).Results Quantitative RQ-PCR demonstrated that the expression of FasL mRNA in sperm from the varicocele group was higher than in the control group. Also, sperm concentration was higher in the controls, when compared to the varicocele group. When submitted to correlation analysis, adolescents with varicocele presented a correlation between sperm concentration and FasL gene expression levels (r=−0.470), not observed in controls. Conclusion Our results allow us to conclude that, in adolescents with varicocele presenting lower sperm concentration, FasL mRNA levels are higher than in adolescents without varicocele.

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“…This laboratory's earliest foray into sperm RNA was in its assessment as a convenient and sensitive marker of successful (or unsuccessful) human vasectomy [55]. We also reasoned that if ejaculate spermatozoa could provide equivalent information on what underlies a nonobstructive subfertile or infertile phenotype, a noninvasively obtained semen sample was surely a preferable and more widely acceptable option to testis biopsy.…”

Section: Historical Contextmentioning

confidence: 99%

Sperm RNA as a Mediator of Genomic Plasticity

Miller

2014

Advances in Biology

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Sperm RNA has been linked recently to trans-generational, non-Mendelian patterns of inheritance. Originally dismissed as "residual" to spermatogenesis, some sperm RNA may have postfertilization functions including the transmission of acquired characteristics. Sperm RNA may help explain how trans-generational effects are transmitted and it may also have implications for assisted reproductive technologies (ART) where sperm are subjected to considerable, ex vivo manual handling. The presence of sperm RNA was originally a controversial topic because nuclear gene expression is switched off in the mature mammalian spermatozoon. With the recent application of next generation sequencing (NGS), an unexpectedly rich and complex repertoire of RNAs has been revealed in the sperm of several species that makes its residual presence counterintuitive. What follows is a personal survey of the science behind our understanding of sperm RNA and its functional significance based on experimental observations from my laboratory as well as many others who have contributed to the field over the years and are continuing to contribute today. The narrative begins with a historical perspective and ends with some educated speculation on where research into sperm RNA is likely to lead us in the next 10 years or so.

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Differential RNA fingerprinting as a tool in the analysis of spermatozoal gene expression

Cited by 86 publications

References 0 publications

Sperm transcriptome profiling in oligozoospermia

Sperm transcriptome profiling in oligozoospermia

Expression of the Fas-ligand gene in ejaculated sperm from adolescents with and without varicocele

Sperm RNA as a Mediator of Genomic Plasticity

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